Objective To explore the mechanism of ZMS regulation of M_2 muscarinic receptor mRNA expression. Methods In vitro cultured CHOm2 cells were divided into ZMS 1 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h), ZMS 2 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h and 1 μg/mL cycloheximide for 12 h) and ZMS 3 group (treatment with 1 μg/mL cycloheximide for 4 h and 1 × 10 ~(-5) mol/L ZMS for 24 h), and their corresponding control groups were also established (substitution of ZMS by DMSO). Actinomycin D was added to cultured CHOm2 cells of each group to inhibit the synthesis of mRNA. CHOm2 cell samples were taken at different time points, the relative expression of M_2 receptor mRNA was detected by Real-time PCR, and half life of M_2 receptor mRNA was calculated. Results Compared with corresponding control groups, the half life of M_2 receptor mRNA of CHOm2 cells in ZMS 1 group and ZMS 2 group was significantly prolonged [(4.75h± 0.54) h vs (2.13 ±0.23) h, P<0.05; (5.43 ±1.13) h vs (2.46 ±0.09) h, P<0.05]. There was no significant difference in half life of M_2 receptor mRNA of CHOm2 cells between ZMS 3 group and its corresponding control [ ( 3.06 ±0.23) h vs (3.00 ± 0.20) h, P > 0.05]. Conclusion De novo protein synthesis is required for the enhancement of M_2 receptor mRNA stability regulated by ZMS.

Objective To study the effect of Neuregulin-1 (NRG) on rat heart failure(HF) after myocardial infarction,and to investigate the underlying mechanism involving in NRG-mediated cardioprotection.Methods 60 adult Wister rats underwent sham operation (n=12) or coronary ligation (n=48) to induce HF.Four weeks after ligation,28 animals with HF were randomly divided into NRG group (rats received rhNRG-1 10 μg · kg-1 · d-1 intravenously for 10 days) or HF group (rats received an equal volume of water intravenously for 10 days).Left ventricular function was detected by echocardiography.Mitochondrial ultrastructure in non-infarcted myocardium was observed by transmission electronic microscopy.Cell apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay.Mitochondrial membrane potential was measured by immunofluorescence method.Caspase-3 activity was determined by commercial kit.The expression of cytochrome C protein in cytoplasm was detected by Western blot.Results Compared with HF group,the left ventricular end-systolic diameter was decreased,and left ventricular ejection fraction and fractional shortening were increased in NRG group (P＜0.05 or 0.01).Apoptosis index was reduced inNRG group as compared with HFgroup [(18.1±3.0)％ vs.(11.9±1.4)％,P＜0.01].Mitochondrial membrane potential was increased and the release of cytochrome c in cytoplasm was reduced in NRG group as compared with HF group [(249.8±7.4) mV vs.(222.2±7.5) mV,(0.356±0.024) vs.(0.664±0.085),both P＜0.01].Caspase-3 activity was decreased in NRG group as compared with HF group (P＜0.01).Conclusions NRG attenuats mitochondrial stress and inhibits myocardial cells apoptosis in heart failure rats after myocardial infarction,which may play an important role in the cardioprotection by NRG.

Objective To study the value of spiral CT in diagnosing renal cell carcinoma(RCC) and to evaluate the correlation of CT findings with histopathologie features in RCC. Methods 279 cases with RCC proven by surgery and pathology underwent plain and contrast-enhanced CT examinations. The clinic characteristics and CT findings were analyzed statistically and compared with surgical results. Results 90.32% lesions showed medium or obvious enhancement on contrast-enhanced CT,with the exception of granule cells for enhanced uniform, the other cell type of RCC showed heterogeneous enhancement. 32.26% of tumors was of more or less short-burres (χ~2 = 38.2,P<0.01) ,and the there was significant relationship between pathological signs of short burr and the renal capsule involved by tumors. CT qualitative diagnostic rate was 91.40%. CT staging and pathologic staging were of the correla-tive coefficient of 0. 84, while there was not obviously correlation between CT classification of RCC and cancer cell types(P>0.05). Conclusion Spiral CT can better show characteristics of RCC, that can improve the diagnostic accuracy and staging aility for RCC.

AIM:To check the antiproliferative and apoptosis-inducing effect of Compound Changtai Granule(Radix et Rhizoma ginseng,Semen coicis,Rhizoma curcumae,etc)(CCG)on human colon carcinoma cell line(SW480).METHODS:Methy thiazolyl tetrazolium chromatomerry was used to observe the influence of several concentration of CCG on SW480 cell proliferation cultured in vitro at the various times,and flow cytometry was applied to examining the change in SW480 cell proliferation cycle and apoptosis rate.RESULTS:CCG could successfully prevent SW480 cell from entering G_0/G_1 phase and G_2/M phase,decrease cell numbers in S phase at the same time.CONCLUSION:CCG has an obvious dose-response relationship in the range of 3.51 to 7.81 mg/mL.

Objective The aim of the study is to detect the expression of IGF -1R and IRS-1 in squa-mous cell lung cancer ( SQCLC) and to explore the role in the development of squamous cell lung cancer and clini -cal significance .Methods Specimens from 246 surgical SQCLC and 40 adjacent normal lung tissues were evalu-ated for IGF-1R and IRS-1 expression by immunohistochemistry .We explored the correlation between IGF -1R and IRS-1,their relationship with clinicopathological parameters and their impact on outcome in SQCLC pa -tients.Results SQCLC of IGF -1R positive expression rate was 54.07%,which was higher than in adjacent normal tissue(32.5%),SQCLC of IRS-1 positive expression rate was 38.21%,which was lower than in adja-cent normal tissue(70%),the pairwise difference was statistically significant (P<0.05);Expression of IGF-1R was correlated with lymph node metastasis (P<0.05),IRS-1 expression was related with degree of differentia-tion and lymph node metastasis (P<0.05).Survival of IGF -1R expression group of patients was significantly shorter than the expression of IGF -1R negative patients ,the survival of the IRS-1 expression was significantly longer than patients who negatively expressed IRS -1,the difference was statistically significant (P=<0.001 and=0.021).IGF-1R was negatively correlated with IRS -1 expression in SQCLC(r=-0.125,P<0.001). Conclusion IGF-1R,IRS-1 are involved in the occurrence of SQCLC ,the development .The IGF-1R is an independent prognostic factor in SQCLC .

Objective To explore the relationship between brachial ankle pulse wave velocity (BaPWV) change and carotid artery plaque formation in middle-aged and elderly patients with essential hypertension(EH).Methods A cross sectional study was conducted in 409 patients who were admitted in Hypertension Department in our hospital from January to September 2013.Their age ranged from 35 to 75 years,with mean aged(53.2±15.0) years.Parallel carotid artery ultrasound and BaPWV examination were performed in all patients.According to whether carotid plaques were present,patients were divided into two groups:carotid plaque group and control group.And carotid plaque group was sub-grouped into normal BaPWV group (BaPWV ＜ 1400cm/s) and increased BaPWV group(BaPWV≥1400 cm/s) according to BaPWV levels.The detection rate of carotid plaque was compared between normal BaPWV group and increased BaPWV group.The correlation between BaPWV and carotid plaques was analyzed by multivariate logistic regression analysis.Results Compared with the control group,carotid plaque group showed the prevalence of carotid plaques was increased along with the increases of age(t=11.0,P=11.0),systolic blood pressure (t=3.87,P=3.87),diastolic blood pressure(t=3.70,P=0.00),pulse pressure(t=6.13,P 6.13),total cholesterol levels(t=2.57,P=0.01).The detection rate of carotid plaque was higher in increased BaPWV group than in normal BaPWV group [62.6％(159/254) vs.43.2％(67/155),x2=14.61,P=0.00].After adjusting for age,blood pressure and other cardiovascular risk factors,the multivariate logistic regression analysis showed that increased BaPWV was an independent risk factor for carotid plaques(OR=2.06,P=0.05),and age,smoking,systolic pressure,diastolic pressure,pulse pressure,total cholesterol,low-density lipoprotein cholesterol,triglyceride levels were positively correlated with carotid plaques.Conclusions BaPWV is one of the independent impact factors for carotid plaques,which plays an important role in early diagnosis and screening for subclinical vascular lesions.

Objective To investigate the safety and efficacy of CO2 fractional laser combined with 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (ALA-PDT) for the treatment of refractory flat wart.Methods The 80 patients with facial flat wart were randomly divided into observation group (40 cases) and control group (40 cases).The observation group was treated with CO2 fractional laser combined with 5-ALA-PDT,and the control group was treated with 5-ALA-PDT only.Those with residual lesions after the first treatment were treated with the second or third times.The interval duration between treatments was 2 weeks.The efficacy and the adverse reaction were evaluated after the 2-week,4-week and 6-month after the last treatment.Results Compared with prior treatment,the facial lesions achieved complete response and improvement.The total effective rate of the observation group were 90.0％ (36/40) and 72.5％ (29/40) for the control group,the difference was statistically significant (P＜0.05).There were 2 cases of pigmentation occurred in observation group and 3 cases in control group.The complications including infection and hypertrophic scar were seldom occurred in the two groups.Conclusions The safety and efficacy of CO2 fractional laser combined with ALA-PDT are better than that with ALA-PDT only in the treatment of refractory flat wart.

Objective To observe the clinical effect of gonadotropin releasing hormone analogues in treatment of the girls with idiopathic central precocious puberty(ICPP). Methods Twelve girls with ICPP received therapy with GnRH-A.Before and after the end of treatment for 6 months,secondary sexual characteristics and growth velocity were observed,pelvic ultrasonic examination,X-ray bone age(BA),serum level of E_2 and LHRH stimulating test were detected,and the predicted adult height(PAH) was analysed. Results After treatment,the size of breasts,uterus and ovaries were significantly reduced.The basic and peak levels of serum LH and FSH by LHRH stimulation were also significantly reduced.The ratio of BA/CA(chronology age)was decreased.The PAH was increased. Conclusions GnRH-A can effectively depress the sexual characteristics,reduce the maturation of BA,and also improve the PAH in girls with ICPP.

AIM:To check the antiproliferative and apoptosis-inducing effect of Compound Changtai Granule(Radix et Rhizoma ginseng,Semen coicis,Rhizoma curcumae,etc)(CCG) on human colon carcinoma cell line(SW480).METHODS:Methy thiazolyl tetrazolium chromatomerry was used to observe the influence of several concentration of CCG on SW480 cell proliferation cultured in vitro at the various times,and flow cytometry was applied to examining the change in SW480 cell proliferation cycle and apoptosis rate.RESULTS:CCG could successfully prevent SW480 cell from entering G0/G1 phase and G2/M phase,decrease cell numbers in S phase at the same time.CONCLUSION:CCG has an obvious dose-response relationship in the range of 3.51 to 7.81 mg/mL.

Objective:To detect the engraftment capability and hematopoietic potential of the expanded cells,establish an optimized project of ex vivo expansion and the SCID mice model of UCB transplantation.Methods:Human cord blood CD34 + cells were clutured in strom free liquid culture system,the sublethally irradiated SCID mice were injected with human expanded cord blood cells by tail vein.Four weeks after inocuration,BM cells of the survived animals are obtained from both femurs and tibias and assessed for the presence of human cells by immunofluorescence staining and PCR.Results:The cells and CFUs of group FL+TPO+SCF+IL 6 were increased significantly,and maintained some proportional CD34 + cells.The CFU of group SCF+IL 6+IL 3+GM CSF+EPO had reduced in week 2,CFC and CD34 + cells and Alu gene were detected in SCID mice BM.Conclusion:The cytokine combination with FL+TPO+SCF+IL 6 can effectively expand UCB CD34 + cells,and the expanded cells can engraft the BM of sublethally irradiated SCID mice and reconstitute hematopoiesis.